具体实施方式
以下参照参考例和实施例进一步说明本发明,但本发明不限于这些例子。
β-内酰胺的制备如(Scheme 4)所示,根据文献(Ojima I. et al.,
Chirality, 2000, 12, 431-441;Ojima I.et al., Bioorg. Med. Chem. 2003,
11, 2867-2888等)报道的合成方法,将末端取代的异丁烯醛与取代的苯胺在脱水剂作用下形成烯夫碱Ⅵ -1-1,未经纯化直接与α-酰氧基乙酰卤在有机碱催化下发生环加成反应获得外消旋的β-内酰胺中间体Ⅵ -1-2, 未经纯化直接进行皂化反应得到中间体Ⅵ -1-3粗品,通过一步重结晶纯化处理得到高纯度的化合物Ⅵ -1-3,三步总收率稳定在70%左右,然后将其进行硅醚化保护,以定量的收率得到化合物Ⅵ -1-4,无需纯化直接于低温下氧化脱除β-内酰胺氨基保护基,经过快速柱层析纯化处理后,以良好的收率得到化合物Ⅵ -1-6, 然后与(Boc)2O、有机碱及催化剂DMAP的作用下进行氨基的Boc的保护,以定量的收率关键中间体Ⅵ -1-10。
卤代的β-内酰胺的制备可根据文献(Ojima I. et al., J. Nat. Prod. 2009, 72, 554–565等)报道的合成方法来完成,将硅醚保护的中间体Ⅵ -1-4进行臭氧氧化断裂双键,以定量的收率得到含有醛基的β-内酰胺中间体Ⅵ -1-5,然后与二溴二氟甲烷/三(二甲氨基)膦(HMPA)或二氟氯乙酸钠/三苯基膦进行Wittig反应,得到二氟取代的内酰胺环Ⅵ -1-7,接着氧化脱除β-内酰胺氨基保护基,以较好的收率得到中间体Ⅵ -1-8, 然后与(Boc)2O)、有机碱及催化剂DMAP的作用下,进行氨基的Boc的保护,以定量的收率关键中间体Ⅵ -1-9。
新型紫杉烷类衍生物的制备可以参照以下方法制备,但本发明不限于这些实施例:
所用试剂:4-二甲氨基吡啶(DMAP)、叔丁基二甲基氯硅烷(TBSCl)、三氟甲磺酸酐(Tf2O)、四丁基氟化铵(TBAF)、硝酸铈铵(CAN)等均为市售试剂,直接用于反应。10-去乙酰巴卡亭(10-DAB)从西安昊轩生物科技有限公司购得。所用石油醚为60~90 oC沸程。未经特殊说明,其它均为普通国产分析纯试剂。其中CH2Cl2经CaH2回流重蒸处理,四氢呋喃(THF)经钠丝重整处理,DMF、吡啶、乙腈、MeOH经3 Å分子筛处理后重蒸。
以薄层色谱(TLC)检测反应进程,所用薄板为烟台化学工业研究所产薄层色谱硅胶预制板(硅胶粒度 10~40m)。紫外检测波长为254 nm;以5%茴香醛-5%浓硫酸-1%冰醋酸-乙醇溶液加热显色。柱层析所用硅胶购自青岛海洋化工厂分厂,规格为100~200目、200~300和300~400目。
生物实验材料:KB、KB/VCR、MCF-7、MCF-7/ADR四种细胞株均为中科院上海药物所药理实验室提供;丙酮酸钠、谷氨酰胺、磺酰罗丹明B (sulforhodamin B,SRB)、二甲基亚砜(DMSO) (均为美国Sigma公司产品);小牛血清(FBS)、DMEM培养基、MEM培养基干粉和胰蛋白酶(均为美国GIBCO公司产品);三氯乙酸、醋酸等其他试剂均为分析纯(国药集团化学试剂有限公司);水为注射用生理盐水。
主要实验仪器:核磁共振波谱仪(JEOL-ECP-600, 内标: TMS、DSS),傅立叶变换红外光谱仪(Nicolet Nexus-470),部分核磁数据来自中科院青岛海洋所500 MHz、复旦大学药学院400 MHz核磁共振波谱仪。细胞计数仪(Beckman 6605698),光吸收酶标仪 (MY190)。
13-
羰基
-7-
脱氧
-7
β
, 8
β
-
亚甲基
-
巴卡亭
III (
Ⅴ
-2):
将化合物7-脱氧-7β, 8β-亚甲基-巴卡亭 III (Ⅴ -1) (2.8 g, 5.0 mmol) 溶于干燥的二氯甲烷(120 mL)中,室温下加入重铬酸吡啶盐(PDC) (2.2 g, 6.0 mmol) 和 4 Å 分子筛(6.0 g),反应3 h,TLC检测无原料剩余后,将混合物抽滤,并反复用乙酸乙酯淋洗数次,将有机层收集,依次用1 M 盐酸(100 mL × 3)、饱和碳酸氢钠水溶液(50 mL × 3)、饱和氯化钠水溶液(50 mL × 3)洗涤,无水硫酸钠干燥,浓缩得到红棕色固体,用乙酸乙酯/石油醚重结晶得到白色粉末状固体13-羰基-7-脱氧-7β, 8β-亚甲基-巴卡亭 III (Ⅴ -2) (2.8 g, 96% yield): 1H
NMR (400 MHz, CDCl3) δ 8.09 (d, J = 7.8 Hz, 2 H, Ph-H),
7.64 (t, J = 7.4 Hz, 1 H, Ph-H), 7.50 (t, J = 7.8 Hz, 2 H, Ph-H), 6.42 (s, 1 H,
H-10), 5.73 (d, J = 7.8 Hz, 1 H, H-2), 4.72 (d, J = 3.9 Hz, 1 H, H-5), 4.31 (d,
J = 8.6 Hz, 1 H, H-20a), 4.19 (d, J = 7.4 Hz, 1 H, H-3), 4.00 (d, J = 8.6 Hz, 1
H, H-20b), 3.06 (d, J = 20.0 Hz, 1 H, H-14a), 2.68 (d, J = 20.0 Hz, 1 H,
H-14b), 2.45 (dt, J = 16.4, 4.3 Hz, 1 H, H-6a), 2.25 (s, 3 H, CH 3CO
in C-10), 2.20-2.31 (m, 1 H, H-19a), 2.13 (s, 3 H, CH 3CO in
C-4), 2.04-2.12 (m, 1 H, H-6b), 2.02 (s, 3 H, H-18), 1.63-1.67 (m, 1 H, H-19b),
1.34 (m, 1 H, H-7), 1.29 (s, 3 H, H-16), 1.26 (s, 3 H, H-17)。
去苯甲酰基
-13-
羰基
-7-
脱氧
-7
β
, 8
β
-
亚甲基
-
巴卡亭
III (
Ⅴ
-3):
将化合物13-羰基-7-脱氧-7β, 8β-亚甲基-巴卡亭 III (Ⅴ -2) (1.1 g, 2.0 mmol) 溶于四氢呋喃(30 mL),加入四丁基氟化铵(1.6 g, 6.0 mmol),室温搅拌6 h,TLC检测无原料剩余后,将反应液浓缩,所得物用乙酸乙酯(150 mL)稀释,依次用1 M盐酸(100 mL×3)、饱和碳酸氢钠水溶液(50 mL × 3)、饱和氯化钠水溶液(50 mL × 3)洗涤,无水硫酸钠干燥,浓缩得到红棕色固体,粗产物经柱层析分离(石油醚/乙酸乙酯 = 1/1)得到橙黄色固体2-去苯甲酰基-13-羰基-7-脱氧-7β, 8β-亚甲基-巴卡亭 III (Ⅴ -3) (672.7 mg, 80% yield):
1H NMR (400 MHz, CDCl3) δ 6.36 (s, 1 H, H-10), 4.73 (d, J =
3.5 Hz, 1 H, H-5), 4.59 (d, J = 9.4 Hz, 1 H, H-20a), 4.50 (d, J = 9.8 Hz, 1 H,
H-20b), 4.08 (d, J = 7.4 Hz, 1 H, H-3), 3.74 (d, J = 7.4 Hz, 1 H, H-2), 2.91
(d, J = 19.6 Hz, 1 H H-14a), 2.61 (dd, J = 19.6, 1.2 Hz, 1 H, H-14b), 2.43 (dt,
J = 16.4, 4.3 Hz, 1 H, H-6a), 2.22 (s, 3 H, CH 3CO in C-4),
2.18 (dd, J = 9.8, 5.5 Hz, 1 H, H-19a), 2.11 (broad d, J = 16.0 Hz, 1 H, H-6b),
1.98 (s, 3 H, CH 3CO in C-10), 1.94 (s, 3 H, H-18), 1.71-1.75
(m, 1 H, H-19b), 1.29-1.33 (m, 1 H, H-7), 1.26 (s, 3 H, H-16), 1.19 (s, 3 H,
H-17)。
去苯甲酰基
-2-(
间
-
三氟甲基苯甲酰基
)-13-
羰基
-7-
脱氧
-7
β
, 8
β
-
亚甲基
-
巴卡亭
III (
Ⅴ
-4a):
将制备的中间体2-去苯甲酰基-13-羰基-7-脱氧-7β, 8β-亚甲基-巴卡亭 III (Ⅴ -3) (84.0 mg, 0.2 mmol),间-三氟甲基苯甲酸 (304.2 mg, 1.0 mmol),和DMAP (24.4 mg, 0.2 mmol)溶于二氯甲烷(20 mL),加入缩合剂二环己基碳二亚胺(DCC) (330.1 mg, 1.6 mmol),于40 °C下搅拌48 h. 将反应液抽滤,有机层用二氯甲烷稀释,依次用1 M 盐酸(10 mL × 3)、饱和碳酸氢钠水溶液(10 mL × 3)、饱和氯化钠水溶液(10 mL × 3)洗涤,无水硫酸钠干燥,浓缩得到白色固体,粗产物经柱层析分离(石油醚/乙酸乙酯 = 1/1)得到白色粉末状固体2-去苯甲酰基-2-(间-三氟甲基苯甲酰基)-13-羰基-7-脱氧-7β, 8β-亚甲基-巴卡亭 III(Ⅴ -4a)(115.8 mg, 91%): 1H
NMR (400 MHz, CDCl3) δ 8.42 (s, 1 H, Ph-H), 8.28 (d, J =
7.8 Hz, 1 H, Ph-H), 7.90 (d, J = 7.8 Hz, 1 H, Ph-H), 7.67 (t, J = 7.8 Hz, 1 H,
Ph-H), 6.43 (s, 1 H, H-10), 5.71 (d, J = 7.4 Hz, 1 H, H-2), 4.75 (d, J = 3.9
Hz, 1 H, H-5), 4.26 (d, J = 8.6 Hz, 1 H, H-20a), 4.22 (d, J = 7.8 Hz, 1 H, H-3),
4.00 (d, J = 8.6 Hz, 1 H, H-20b), 3.00 (d, J = 20.0 Hz, 1 H, H-14a), 2.69 (d, J
= 20.0 Hz, 1 H, H-14b), 2.46 (dt, J = 16.0, 3.9 Hz, 1 H, H-6a), 2.30 (dd, J =
10.2, 5.1 Hz, 1 H, H-19a), 2.26 (s, 3 H, CH 3CO in C-10), 2.13
(s, 3 H, CH 3CO in C-4), 2.09 (broad s, 1 H, H-6b), 2.03 (s, 3
H, H-18), 1.67 (t like, J = 6.7 Hz, 1 H, H-19b), 1.35 (m, 1 H, H-7), 1.30 (s, 3
H, H-16), 1.25 (s, 3 H, H-17)。
根据相同的方法可以制备其他的C-2苯甲酰基被取代的中间体(Ⅴ -4b~ Ⅴ -4h):
2- 去苯甲酰基 -2-( 间 - 甲基苯甲酰基 )-13- 羰基 -7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -4b): 1H NMR (400
MHz, CDCl3) δ 7.88-7.92 (m, 2 H, Ph-H), 7.36-7.45 (m, 2 H, Ph-H), 6.42 (s, 1 H, H-10),
5.71 (d, J = 7.8 Hz, 1 H, H-2), 4.73 (d, J = 3.5 Hz, 1 H, H-5), 4.31 (d, J =
8.6 Hz, 1 H, H-20a), 4.19 (d, J = 7.4 Hz, 1 H, H-3), 3.99 (d, J = 9.0 Hz, 1 H,
H-20b), 3.06 (d, J = 20.0 Hz, 1 H, H-14a), 2.69 (d, J = 20.0 Hz, 1 H, H-14b),
2.41-2.47 (m, 4 H, H-6a and PhCH 3), 2.27-2.31 (m, 1 H,
H-19a), 2.25 (s, 3 H, CH 3CO in C-10), 2.13 (s, 3 H, CH 3CO
in C-4), 2.08 (broad s, 1 H, H-6b), 2.03 (s, 3 H, H-18), 1.66 (t like, J = 6.3
Hz, 1 H, H-19b), 1.33 (m, 1 H, H-7), 1.30 (s, 3 H, H-16), 1.25 (s, 3 H, H-17)。
去苯甲酰基 -2-( 间 - 甲氧基苯甲酰基 )-13- 羰基 -7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -4c): 1H NMR (400
MHz, CDCl3) δ 7.70 (d, J = 7.8 Hz, 1 H, Ph-H), 7.62 (s, 1 H, Ph-H), 7.40 (t, J = 8.2
Hz, 1 H, Ph-H), 7.17 (d, J = 8.2 Hz, 1 H, Ph-H), 6.43 (s, 1 H, H-10), 5.73 (d,
J = 7.4 Hz, 1 H, H-2), 4.73 (d, J = 3.5 Hz, 1 H, H-5), 4.35 (d, J = 8.6 Hz, 1
H, H-20a), 4.20 (d, J = 7.4 Hz, 1 H, H-3), 4.00 (d, J = 8.6 Hz, 1 H, H-20b),
3.87 (s, 3 H, OCH 3), 3.05 (d, J = 20.0 Hz, 1 H, H-14a), 2.69
(d, J = 20.0 Hz, 1 H, H-14b), 2.46 (dt, J = 16.0, 4.3 Hz, 1 H, H-6a), 2.23-2.31
(m, J = 10.2, 5.1 Hz, 1 H, H-19a), 2.26 (s, 3 H, CH 3CO in
C-10), 2.12 (s, 3 H, CH 3CO in C-4), 2.08-2.13 (m, 1 H, H-6b),
2.03 (s, 3 H, H-18), 1.67 (t like, J = 6.7 Hz, 1 H, H-19b), 1.28-1.33 (m, 1 H,
H-7), 1.30 (s, 3 H, H-16), 1.25 (s, 3 H, H-17)。
去苯甲酰基 -2-( 间 - 三氟甲氧基苯甲酰基 )-13- 羰基 -7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -4d): 1H NMR (400
MHz, CDCl3) δ 8.04 (d, J = 7.8 Hz, 1 H, Ph-H), 7.97 (s, 1 H, Ph-H), 7.54-7.58 (m, 1 H,
Ph-H), 7.49 (t, J = 7.8 Hz, 1 H, Ph-H), 6.43 (s, 1 H, H-10), 5.72 (d, J = 7.8
Hz, 1 H, H-2), 4.74 (broad s, 1 H, H-5), 4.29 (d, J = 8.2 Hz, 1 H, H-20a), 4.21
(d, J = 7.8 Hz, 1 H, H-3), 3098 (d, J = 8.2 Hz, 1 H, H-20b), 3.00 (d, J = 20.0
Hz, 1 H, H-14a), 2.69 (d, J = 20.0 Hz, 1 H, H-14b), 2.46 (dt, J = 16.0, 4.3 Hz,
1 H, H-6a), 2.28-2.30 (m, 1 H, H-19a), 2.26 (s, 3 H, CH 3CO in
C-10), 2.12 (s, 3 H, CH 3CO in C-4), 2.09 (broad s, 1 H,
H-6b), 2.03 (s, 3 H, H-18), 1.66 (t like, J = 7.0 Hz, 1 H, H-19b), 1.35 (m, 1
H, H-7), 1.29 (s, 3 H, H-16), 1.25 (s, 3 H, H-17)。
去苯甲酰基 -2-( 间 - 氟苯甲酰基 )-13- 羰基 -7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -4e): 1H NMR (400
MHz, CDCl3) δ 7.89 (d, J = 7.8 Hz, 1 H, Ph-H), 7.78 (dd, J = 9.0, 1.5 Hz, 1 H, Ph-H),
7.46-7.52 (m, 1 H, Ph-H), 7.32-7.37 (m, 1 H, Ph-H), 6.43 (s, 1 H, H-10), 5.71
(d, J = 7.4 Hz, 1 H, H-2), 4.74 (d, J = 3.5 Hz, 1 H, H-5), 4.31 (d, J = 8.6 Hz,
1 H, H-20a), 4.20 (d, J = 7.8 Hz, 1 H, H-3), 3.99 (d, J = 8.6 Hz, 1 H, H-20b),
3.02 (d, J = 20.0 Hz, 1 H, H-14a), 2.69 (d, J = 20.0 Hz, 1 H, H-14b), 2.46 (dt,
J = 16.0, 4.3 Hz, 1 H, H-6a), 2.30 (dd, J = 9.8, 5.9 Hz, 1 H, H-19a), 2.25 (s,
3 H, CH 3CO in C-10), 2.14 (s, 3 H, CH 3CO in
C-4), 2.09 (broad s, 1 H, H-6b), 2.03 (s, 3 H, H-18), 1.66 (t like, J = 6.7 Hz,
1 H, H-19b), 1.35 (m, 1 H, H-7), 1.30 (s, 3 H, H-16), 1.25 (s, 3 H, H-17)。
去苯甲酰基 -2-( 间 - 氯苯甲酰基 )-13- 羰基 -7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -4f): 1H NMR (400
MHz, CDCl3) δ 8.10 (s, 1 H, Ph-H), 7.98 (d, J = 7.8 Hz, 1 H, Ph-H), 7.61 (d, J = 7.8
Hz, 1 H, Ph-H), 7.45 (t, J = 7.8 Hz, 1 H, Ph-H), 6.42 (s, 1 H, H-10), 5.69 (d,
J = 7.4 Hz, 1 H, H-2), 4.74 (d, J = 3.9 Hz, 1 H, H-5), 4.30 (d, J = 8.6 Hz, 1
H, H-20a), 4.21 (d, J = 7.4 Hz, 1 H, H-3), 3.98 (d, J = 8.6 Hz, 1 H, H-20b),
3.02 (d, J = 20.0 Hz, 1 H, H-14a), 2.69 (d, J = 20.3 Hz, 1 H, H-14b), 2.45 (dt,
J = 16.0, 5.1 Hz, 1 H, H-6a), 2.29 (dd, J = 10.2, 5.1 Hz, 1 H, H-19a), 2.26 (s,
3 H, CH 3CO in C-10), 2.15 (s, 3 H, CH 3CO in
C-4), 2.11 (broad s, 1 H, H-6b), 2.03 (s, 3 H, H-18), 1.66 (t like, J = 6.7 Hz,
1 H, H-19b), 1.34 (m, 1 H, H-7), 1.29 (s, 3 H, H-16), 1.25 (s, 3 H, H-17)。
去苯甲酰基 -2-( 间 - 氰基苯甲酰基 )-13- 羰基 -7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -4g): 1H NMR (400
MHz, CDCl3) δ 8.40 (s, 1 H, Ph-H), 8.31 (d, J = 7.8 Hz, 1 H, Ph-H), 7.92 (d, J = 7.4
Hz, 1 H, Ph-H), 7.67 (t, J = 7.8 Hz, 1 H, Ph-H), 6.43 (s, 1 H, H-10), 5.71 (d,
J = 7.4 Hz, 1 H, H-2), 4.74 (d, J = 3.5 Hz, 1 H, H-5), 4.24 (d, J = 8.6 Hz, 1
H, H-20a), 4.22 (d, J = 7.8 Hz, 1 H, H-3), 3.98 (d, J = 8.6 Hz, 1 H, H-20b),
2.99 (d, J = 20.0 Hz, 1 H, H-14a), 2.69 (d, J = 20.0 Hz, 1 H, H-14b), 2.46 (dt,
J = 16.0, 4.3 Hz, 1 H, H-6a), 2.30 (dd, J = 10.6, 5.1 Hz, 1 H, H-19a), 2.26 (s,
3 H, CH 3CO in C-10), 2.16 (s, 3 H, CH 3CO in
C-4), 2.12 (d, J = 16.0 Hz, 1 H, H-6b), 2.03 (s, 3 H, H-18), 1.66 (t like, J =
6.7 Hz, 1 H, H-19b), 1.34 (m, 1 H, H-7), 1.30 (s, 3 H, H-16), 1.25 (s, 3 H,
H-17)。
去苯甲酰基 -2-( 间 - 叠氮基苯甲酰基 )-13- 羰基 -7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -4h): 1H NMR (400
MHz, CDCl3) δ 7.87 (d, J = 7.8 Hz, 1 H, Ph-H), 7.77 (s, 1 H, Ph-H), 7.49 (t, J = 8.2
Hz, 1 H, Ph-H), 7.28 (d, J = 8.2 Hz, 1 H, Ph-H), 6.43 (s, 1 H, H-10), 5.73 (d,
J = 7.4 Hz, 1 H, H-2), 4.74 (d, J = 3.9 Hz, 1 H, H-5), 4.32 (d, J = 8.6 Hz, 1
H, H-20a), 4.21 (d, J = 7.4 Hz, 1 H, H-3), 3.99 (d, J = 8.6 Hz, 1 H, H-20b),
3.02 (d, J = 20.0 Hz, 1 H, H-14a), 2.69 (d, J = 20.0 Hz, 1 H, H-14b), 2.46 (dt,
J = 16.4 Hz, 1 H, H-6a), 2.28-2.32 (m, 1 H, H-19a), 2.26 (s, 3 H, CH 3CO
in C-10), 2.14 (s, 3 H, CH 3CO in C-4), 2.09 (broad s, 1 H,
H-6b), 2.03 (s, 3 H, H-18), 1.66 (t like, J = 6.7 Hz, 1 H, H-19b), 1.35 (m, 1
H, H-7), 1.30 (s, 3 H, H-16), 1.25 (s, 3 H, H-17)。
去苯甲酰基
-2-(
间
-
三氟甲基苯甲酰基
)-7-
脱氧
-7
β
, 8
β
-
亚甲基
-
巴卡亭
III (
Ⅴ
-5a):
将制备的中间体2-去苯甲酰基-2-(间-三氟甲基苯甲酰基)-13-羰基-7-脱氧-7β, 8β-亚甲基-巴卡亭 III (Ⅴ -4a) (102.0 mg, 0.16 mmol)溶于甲醇(2 mL)和四氢呋喃(10 mL),冰浴下加入硼氢化钠(121.1 mg, 3.2 mmol),室温反应5 h后,加入饱和氯化铵(10 mL)终止反应,将混合物用乙酸乙酯(30 mL × 3)萃取,合并有机相,依次用1 M盐酸(10 mL × 3)、饱和碳酸氢钠水溶液(10 mL × 3)、饱和氯化钠水溶液(10 mL×3)洗涤,无水硫酸钠干燥,浓缩得到白色固体,粗产物经柱层析分离(石油醚/乙酸乙酯 = 2/1)得到白色粉末状固体2-去苯甲酰基-2-(间-三氟甲基苯甲酰基)-7-脱氧-7β, 8β-亚甲基-巴卡亭 III (Ⅴ -5a) (100.2 mg, 98% yield):
1H NMR (400 MHz, CDCl3) δ 8.47 (s, 1 H, Ph-H), 8.31 (d, J = 7.8
Hz, 1 H, Ph-H), 7.88 (d, J = 7.4 Hz, 1 H, Ph-H), 7.65 (t, J = 7.8 Hz, 1 H,
Ph-H), 6.35 (s, 1 H, H-10), 5.61 (d, J = 7.8 Hz, 1 H, H-2), 4.83 (t, J = 7.4
Hz, 1 H, H-13), 4.77 (d, J = 3.5 Hz, 1 H, H-5), 4.25 (d, J = 8.2 Hz, 1 H,
H-20a), 4.20 (d, J = 7.4 Hz, 1 H, H-3), 4.02 (d, J = 8.2 Hz, 1 H, H-20b), 2.48
(dt, J = 16.4, 4.3 Hz, 1 H, H-6a), 2.23-2.35 (m, 3 H, H-14 and H-19a), 2.25 (s,
3 H, CH 3CO in C-10), 2.21 (s, 3 H, CH 3CO in
C-4), 2.10 (d, J = 16.0 Hz, 1 H, H-6b), 2.03 (s, 3 H, H-18), 1.63-1.67 (m, 1 H,
H-19b), 1.35 (m, 1 H, H-7), 1.22 (s, 3 H, H-16), 1.10 (s, 3 H, H-17)。
根据相同的方法可以制备其他的C-2苯甲酰基被取代的中间体(Ⅴ -5b~ Ⅴ -5h):
2- 去苯甲酰基 -2-( 间 - 甲基苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -5b): 1H NMR (400
MHz, CDCl3) δ 7.93-7.97 (m, 2 H, Ph-H), 7.43 (d, J = 7.8 Hz, 1 H, Ph-H), 7.37 (t, J =
7.3 Hz, 1 H, Ph-H), 6.35 (s, 1 H, H-10), 5.61 (d, J = 7.8 Hz, 1 H, H-2), 4.84
(t, J = 7.8 Hz, 1 H, H-13), 4.76 (d, J = 3.4 Hz, 1 H, H-5), 4.32 (d, J = 8.8
Hz, 1 H, H-20a), 4.18 (d, J = 7.8 Hz, 1 H, H-3), 4.03 (d, J = 8.8 Hz, 1 H,
H-20b), 2.48 (dt, J = 16.1 Hz, 1 H, H-6a), 2.23-2.35 (m, 3 H, H-19a), 2.44 (s,
3 H, PhCH 3), 2.27 (s, 3 H, CH 3CO in C-10),
2.23-2.38 (m, 2 H, H-14), 2.22 (s, 3 H, CH 3CO in C-4), 2.10
(d, J = 15.6 Hz, 1 H, H-6b), 2.03 (s, 3 H, H-18), 1.65 (t like, 1 H, H-19b),
1.34 (m, 1 H, H-7), 1.23 (s, 3 H, H-16), 1.10 (s, 3 H, H-17)。
去苯甲酰基 -2-( 间 - 甲氧基苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -5c): 1H NMR (400
MHz, CDCl3) δ 7.73 (d, J = 7.8 Hz, 1 H, Ph-H), 7.66 (d, J = 2.7 Hz, 1 H, Ph-H), 7.39
(t, J = 7.8 Hz, 1 H, Ph-H), 7.15 (dd, J = 8.2, 2.0 Hz, 1 H, Ph-H), 6.34 (s, 1
H, H-10), 5.62 (d, J = 7.8 Hz, 1 H, H-2), 4.83 (t, J = 7.4 Hz, 1 H, H-13), 4.75
(d, J = 3.5 Hz, 1 H, H-5), 4.34 (d, J = 8.6 Hz, 1 H, H-20a), 4.18 (d, J = 7.8
Hz, 1 H, H-3), 4.03 (d, J = 8.6 Hz, 1 H, H-20b), 3.87 (s, 3 H, OCH 3),
2.48 (dt, J = 16.0, 4.3 Hz, 1 H, H-6a), 2.23-2.37 (m, 3 H, H-14 and H-19a),
2.25, 2.20, 2.09 (dd, J = 16.0, 6.3 Hz, 1 H, H-6b), 2.04 (s, 3 H, H-18),
1.63-1.67 (m, 1 H, H-19b), 1.33 (m, 1 H, H-7), 1.22 (s, 3 H, H-16), 1.10 (s, 3
H, H-17)。
去苯甲酰基 -2-( 间 - 三氟甲氧基苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -5d): 1H NMR (400
MHz, CDCl3) δ 8.07 (d, J = 7.4 Hz, 1 H, Ph-H), 8.02 (s, 1 H, Ph-H), 7.54 (t, J = 7.8
Hz, 1 H, Ph-H), 7.47 (d, J = 7.8 Hz, 1 H, Ph-H), 6.34 (s, 1 H, H-10), 5.60 (d,
J = 7.4 Hz, 1 H, H-2), 4.83 (t, J = 8.2 Hz, 1 H, H-13), 4.76 (d, J = 4.3 Hz, 1
H, H-5), 4.27 (d, J = 8.2 Hz, 1 H, H-20a), 4.19 (d, J = 7.4 Hz, 1 H, H-3), 4.02
(d, J = 8.2 Hz, 1 H, H-20b), 2.48 (dt, J = 16.0, 4.3 Hz, 1 H, H-6a), 2.23-2.36
(m, 3 H, H-14 and H-19a), 2.24 (s, 3 H, CH 3CO in C-10), 2.21
(s, 3 H, CH 3CO in C-4), 2.10 (d, J = 16.0 Hz, 1 H, H-6b),
2.02 (s, 3 H, H-18), 1.64 (m, 1 H, H-19b), 1.34 (m, 1 H, H-7), 1.21 (s, 3 H,
H-16), 1.09 (s, 3 H, H-17)。
去苯甲酰基 -2-( 间 - 氟苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -5e): 1H NMR (400
MHz, CDCl3) δ 8.47 (s, 1 H, Ph-H), 8.31 (d, J = 7.8 Hz, 1 H, Ph-H), 7.88 (d, J = 7.4
Hz, 1 H, Ph-H), 7.65 (t, J = 7.8 Hz, 1 H, Ph-H), 6.35 (s, 1 H, H-10), 5.61 (d,
J = 7.8 Hz, 1 H, H-2), 4.83 (t, J = 7.4 Hz, 1 H, H-13), 4.77 (d, J = 3.5 Hz, 1
H, H-5), 4.25 (d, J = 8.2 Hz, 1 H, H-20a), 4.20 (d, J = 7.4 Hz, 1 H, H-3), 4.02
(d, J = 8.2 Hz, 1 H, H-20b), 2.48 (dt, J = 16.4, 4.3 Hz, 1 H, H-6a), 2.23-2.35
(m, 3 H, H-14 and H-19a), 2.25 (s, 3 H, CH 3CO in C-10), 2.21
(s, 3 H, CH 3CO in C-4), 2.10 (d, J = 16.0 Hz, 1 H, H-6b),
2.03 (s, 3 H, H-18), 1.63-1.67 (m, 1 H, H-19b), 1.35 (m, 1 H, H-7), 1.22 (s, 3
H, H-16), 1.10 (s, 3 H, H-17)。
去苯甲酰基 -2-( 间 - 氯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -5f): 1H NMR (400
MHz, CDCl3) δ 8.15 (s, 1 H, Ph-H), 8.00 (d, J = 7.8 Hz, 1 H, Ph-H), 7.59 (d, J = 7.8
Hz, 1 H, Ph-H), 7.43 (t, J = 7.8 Hz, 1 H, Ph-H), 6.34 (s, 1 H, H-10), 5.57 (d,
J = 7.8 Hz, 1 H, H-2), 4.82 (t, J = 7.4 Hz, 1 H, H-13), 4.76 (d, J = 3.5 Hz, 1
H, H-5), 4.28 (d, J = 8.2 Hz, 1 H, H-20a), 4.18 (d, J = 7.8 Hz, 1 H, H-3), 4.01
(d, J = 8.6 Hz, 1 H, H-20b), 2.48 (dt, J = 16.0, 4.3 Hz, 1 H, H-6a), 2.22-2.34
(m, 3 H, H-14 and H-19a), 2.27 (s, 3 H, CH 3CO in C-10), 2.21
(s, 3 H, CH 3CO in C-4), 2.09 (d, J = 16.0 Hz, 1 H, H-6b),
2.02 (s, 3 H, H-18), 1.62-1.65 (m, 1 H, H-19b), 1.35 (m, 1 H, H-7), 1.21 (s, 3
H, H-16), 1.09 (s, 3 H, H-17)。
去苯甲酰基 -2-( 间 - 氰基苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -5g): 1H NMR (400
MHz, CDCl3) δ 8.45 (s, 1 H, Ph-H), 8.34 (d, J = 7.8 Hz, 1 H, Ph-H), 7.90 (d, J = 7.4
Hz, 1 H, Ph-H), 7.65 (t, J = 7.8 Hz, 1 H, Ph-H), 6.34 (s, 1 H, H-10), 5.59 (d,
J = 7.4 Hz, 1 H, H-2), 4.83 (t, J = 7.4 Hz, 1 H, H-13), 4.76 (broad s, 1 H,
H-5), 4.19-4.23 (m, 2 H, H-20a and H-3), 4.01 (d, J = 8.6 Hz, 1 H, H-20b), 2.48
(dt, J = 16.4, 4.3 Hz, 1 H, H-6a), 2.22-2.35 (m, 3 H, H-14 and H-19a), 2.28 (s,
3 H, CH 3CO in C-10), 2.21 (s, 3 H, CH 3CO in
C-4), 2.10 (d, J = 16.4 Hz, 1 H, H-6b), 2.03 (s, 3 H, H-18), 1.64 (m, 1 H,
H-19b), 1.35 (m, 1 H, H-7), 1.21 (s, 3 H, H-16), 1.10 (s, 3 H, H-17)。
去苯甲酰基 -2-( 间 - 叠氮基苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 - 巴卡亭 III ( Ⅴ -5h): 1H NMR (400
MHz, CDCl3) δ 7.90 (d, J = 7.4 Hz, 1 H, Ph-H), 7.81 (s, 1 H, Ph-H), 7.48 (t, J = 7.8
Hz, 1 H, Ph-H), 7.25 (s, 1 H, Ph-H), 6.34 (s, 1 H, H-10), 5.62 (d, J = 7.8 Hz,
1 H, H-2), 4.83 (t, J = 7.1 Hz, 1 H, H-13), 4.75 (d, J = 3.9 Hz, 1 H, H-5),
4.30 (d, J = 8.6 Hz, 1 H, H-20a), 4.18 (d, J = 7.4 Hz, 1 H, H-3), 4.02 (d, J =
8.6 Hz, 1 H, H-20b), 2.48 (dt, J = 16.0, 4.3 Hz, 1 H, H-6a), 2.23-2.35 (m, 3 H,
H-14 and H-19a), 2.26 (s, 3 H, CH 3CO in C-10), 2.20 (s, 3 H,
CH 3CO in C-4), 2.09 (d, J = 16.0 Hz, 1 H, H-6b), 2.02 (s, 3
H, H-18), 1.62-1.65 (m, 1 H, H-19b), 1.34 (m, 1 H, H-7), 1.21 (s, 3 H, H-16),
1.10 (s, 3 H, H-17)。
去苯甲酰基
-2-(
间
-
三氟甲基苯甲酰基
)-7-
脱氧
-7
β
, 8
β
-
亚甲基
-10-
乙酰氧基
-
多西紫杉醇
(
Ⅰ
-1a)
:
将中间体2-去苯甲酰基-2-(间-三氟甲基苯甲酰基)-7-脱氧-7β, 8β-亚甲基-巴卡亭 III (Ⅴ -5a) (68.0 mg, 0.11 mmol)、(3R, 4S)-1-t-BOC-3-OTES-β-内酰胺(48.5 mg, 0.13 mmol) 溶于干燥的THF(15 mL)中,氩气保护下将反应液冷却至-40 °C,滴加碱LiHMDS (165.0 µL, 0.17
mmol, 1.0 M in THF),继续反应1.5 h,检测反应结束后,加入饱和氯化铵(10 mL)终止反应,将混合物用乙酸乙酯(30 mL × 3)萃取,合并有机相,依次用1 M 盐酸(10 mL × 3)、饱和碳酸氢钠水溶液(10 mL × 3)、饱和氯化钠水溶液(10 mL×3)洗涤,无水硫酸钠干燥,浓缩得到泡沫状固体。将粗产物溶于THF (15 mL),加入TBAF(86.4 mg, 0.33 mmol),40 °C下反应3 h,加入饱和氯化铵(10 mL)终止反应,将混合物用乙酸乙酯(30 mL × 3)萃取,合并有机相,依次用1 M 盐酸(10 mL × 3)、饱和碳酸氢钠水溶液(10 mL × 3)、饱和氯化钠水溶液(10 mL × 3)洗涤,无水硫酸钠干燥,浓缩得到泡沫状固体,粗产物经柱层析分离(石油醚/乙酸乙酯 = 3/1)得到白色粉末状固体2-去苯甲酰基-2-(间-三氟甲基苯甲酰基)-7-脱氧-7β, 8β-亚甲基-10-乙酰氧基-多西紫杉醇(Ⅰ -1a) (88.2 mg, 86% yield for
two steps): 1H NMR (400 MHz, CDCl3) δ 8.47 (s, 1 H, Ph-H), 8.34
(d, J = 7.8 Hz, 1 H, Ph-H), 7.89 (d, J = 7.3 Hz, 1 H, Ph-H), 7.67 (t, J = 7.8
Hz, 1 H, Ph-H), 7.31-7.39 (m, 5 H, Ph-H), 6.33 (s, 1 H, H-10), 6.20 (brt, 1 H,
H-13), 5.64 (d, J = 7.8 Hz, 1 H, H-2), 5.31 (d, J = 8.8 Hz, 1 H, CONH),
5.24 (brd, J = 7.8 Hz, 1 H, H-3′), 4.75 (broad s, 1 H, H-5), 4.59 (s, 1 H, H-2′), 4.27 (d, J = 8.3 Hz, 1
H, H-20a), 4.12 (d, J = 7.8 Hz, 1 H, H-3), 4.02 (d, J = 8.3 Hz, 1 H, H-20b),
2.46 (dt like, J = 16.6, 1 H, H-6a), 2.33 (s, 3 H, CH 3CO in
C-4), 2.24-2.29 (m, 3 H, H-19a and H-14), 2.21 (s, 3 H, CH 3CO
in C-10), 2.11 (d, J = 15.6 Hz, 1 H, H-6b), 1.85 (s, 3 H, CH 3
in C-18), 1.67 (t like, 1 H, H-19b), 1.36 (m, 1 H, H-7), 1.25-1.28 (m, 15 H, Me 3C,
H-16 and H-17)。
根据相同的方法可以制备其他的C-2苯甲酰基被取代的对接产物(Ⅰ -1b ~ Ⅰ -1h):
2- 去苯甲酰基 -2-( 间 - 甲基苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -1b): 1H NMR (400 MHz, CDCl3)
δ 7.98 (s, 1 H, Ph-H), 7.94
(d, J = 5.9 Hz, 1 H, Ph-H), 7.28-7.39 (m, 7 H, Ph-H), 6.32 (s, 1 H, H-10), 6.26
(brt, 1 H, H-13), 5.64 (d, J = 7.0 Hz, 1 H, H-2), 5.36 (broad d, 1 H, CONH),
5.26 (broad d, J = 7.8 Hz, 1 H, H-3′), 4.73 (broad s, 1 H, H-5), 4.61 (s,
1 H, H-2′), 4.31 (d, J = 8.6 Hz, 1
H, H-20a), 4.08 (d, J = 7.0 Hz, 1 H, H-3), 4.02 (d, J = 8.6 Hz, 1 H, H-20b),
3.33 (broad s, 1 H), 2.43-2.47 (m, 1 H, H-6a), 2.43 (s, 1 H, PhCH 3),
2.37 (s, 3 H, CH 3CO in C-4), 2.24-2.26 (m, 3 H, H-19a and
H-14), 2.20 (s, 3 H, CH 3CO in C-10), 2.09 (d, J = 16.0 Hz, 1
H, H-6b), 1.84 (s, 3 H, CH 3 in C-18), 1.66 (t like, 1 H,
H-19b), 1.36 (m, 1 H, H-7), 1.25-1.28 (m, 15 H, Me 3C, H-16
and H-17)。
去苯甲酰基 -2-( 间 - 甲氧基苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -1c): 1H NMR (400 MHz, CDCl3)
δ 7.75 (d, J = 7.8 Hz, 2 H,
Ph-H), 7.74 (s, 1 H, Ph-H), 7.31-7.43 (m, 6 H, Ph-H), 7.14 (d, J = 7.4 Hz, 1 H,
Ph-H), 6.32 (s, 1 H, H-10), 6.26 (brt, 1 H, H-13), 5.66 (d, J = 7.4 Hz, 1 H,
H-2), 5.35 (d, J = 9.0 Hz, 1 H, CONH), 5.28 (brd, J = 7.8 Hz, 1 H, H-3′), 4.73 (broad s, 1 H,
H-5), 4.60 (s, 1 H, H-2′), 4.36 (d, J = 8.6 Hz, 1 H, H-20a), 4.08 (d, J = 7.8 Hz, 1 H, H-3), 4.03
(d, J = 8.6 Hz, 1 H, H-20b), 2.46 (dt, J = 15.3, 4.8 Hz, 1 H, H-6a), 2.36 (s, 3
H, CH 3CO in C-4), 2.20 (s, 3 H, CH 3CO in
C-10), 2.22-2.24 (m, 3 H, H-19a and H-14), 2.10 (d, J = 16.0 Hz, 1 H, H-6b),
1.84 (s, 3 H, CH 3 in C-18), 1.67 (m, 1 H, H-19b), 1.33 (m, 1
H, H-7), 1.26 (m, 15 H, Me 3C, H-16 and H-17)。
去苯甲酰基 -2-( 间 - 三氟甲氧基苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -1d): 1H NMR (400 MHz, CDCl3)
δ 8.47 (s, 1 H, Ph-H), 8.34
(d, J = 7.8 Hz, 1 H, Ph-H), 7.89 (d, J = 7.3 Hz, 1 H, Ph-H), 7.67 (t, J = 7.8
Hz, 1 H, Ph-H), 7.31-7.39 (m, 5 H, Ph-H), 6.33 (s, 1 H, H-10), 6.20 (brt, 1 H,
H-13), 5.64 (d, J = 7.8 Hz, 1 H, H-2), 5.31 (d, J = 8.8 Hz, 1 H, CONH),
5.24 (brd, J = 7.8 Hz, 1 H, H-3′), 4.75 (broad s, 1 H, H-5), 4.59 (s, 1 H, H-2′), 4.27 (d, J = 8.3 Hz, 1
H, H-20a), 4.12 (d, J = 7.8 Hz, 1 H, H-3), 4.02 (d, J = 8.3 Hz, 1 H, H-20b),
2.46 (dt like, J = 16.6, 1 H, H-6a), 2.33 (s, 3 H, CH 3CO in
C-4), 2.24-2.29 (m, 3 H, H-19a and H-14), 2.21 (s, 3 H, CH 3CO
in C-10), 2.11 (d, J = 15.6 Hz, 1 H, H-6b), 1.85 (s, 3 H, CH 3
in C-18), 1.67 (t like, 1 H, H-19b), 1.36 (m, 1 H, H-7), 1.25-1.28 (m, 15 H, Me 3C,
H-16 and H-17)。
去苯甲酰基 -2-( 间 - 氟苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -1e): 1H NMR (400 MHz, CDCl3)
δ 7.95 (d, J = 7.7 Hz, 1 H,
Ph-H), 7.84 (d, J = 9.0 Hz, 1 H, Ph-H), 7.32-7.54 (m, 7 H, Ph-H), 6.32 (s, 1 H,
H-10), 6.25 (brt, 1 H, H-13), 5.63 (d, J = 7.8 Hz, 1 H, H-2), 5.33 (d, J = 8.8
Hz, 1 H, CONH), 5.24 (brd, J = 7.8 Hz, 1 H, H-3′), 4.75 (broad s, 1 H,
H-5), 4.59 (s, 1 H, H-2′), 4.27 (d, J = 8.3 Hz, 1 H, H-20a), 4.12 (d, J = 7.8 Hz, 1 H, H-3), 4.02
(d, J = 8.3 Hz, 1 H, H-20b), 2.46 (dt like, J = 16.6, 1 H, H-6a), 2.33 (s, 3 H,
CH 3CO in C-4), 2.24-2.29 (m, 3 H, H-19a and H-14), 2.21 (s, 3
H, CH 3CO in C-10), 2.11 (d, J = 15.6 Hz, 1 H, H-6b), 1.85 (s,
3 H, CH 3 in C-18), 1.67 (t like, 1 H, H-19b), 1.36 (m, 1 H,
H-7), 1.25-1.28 (m, 15 H, Me 3C, H-16 and H-17);
2- 去苯甲酰基 -2-( 间 - 氯苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -1f): 1H NMR (400 MHz, CDCl3)
δ 8.16 (s, 1 H, Ph-H), 8.04
(d, J = 7.8 Hz, 1 H, Ph-H), 7.72 (dd, J = 5.9, 3.5 Hz, 1 H, Ph-H), 7.56 (d, J =
8.2 Hz, 1 H, Ph-H), 7.53 (dd, J = 5.9, 3.5 Hz, 1 H, Ph-H), 7.46 (t, J = 7.8 Hz,
1 H, Ph-H), 7.30-7.41 (m, 3 H, Ph-H), 6.35 (s, 1 H, H-10), 6.22 (brt, 1 H,
H-13), 5.62 (d, J = 7.4 Hz, 1 H, H-2), 5.33 (d like, 1 H, CONH), 5.27
(brd, 1 H, H-3′), 4.74 (broad s, 1 H,
H-5), 4.59 (s, 1 H, H-2′), 4.30 (d, J = 8.6 Hz, 1 H, H-20a), 4.09 (d, J = 7.8 Hz, 1 H, H-3), 4.01
(d, J = 8.6 Hz, 1 H, H-20b), 2.45 (dt, J = 16.1, 4.3 Hz, 1 H, H-6a), 2.36 (s, 3
H, CH 3CO in C-4), 2.22-2.26 (m, 3 H, H-19a and H-14), 2.21
(s, 3 H, CH 3CO in C-10), 2.14 (d, J = 13.7 Hz, 1 H, H-6b),
1.84 (s, 3 H, CH 3 in C-18), 1.67 (t like, 1 H, H-19b), 1.35
(m, 1 H, H-7), 1.25-1.28 (m, 15 H, Me 3C, H-16 and H-17);
2- 去苯甲酰基 -2-( 间 - 氰基苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -1g): 1H NMR (400 MHz, CDCl3)
δ 8.48 (s, 1 H, Ph-H), 8.38
(d, J = 7.7 Hz, 1 H, Ph-H), 7.88 (d, J = 7.7 Hz, 1 H, Ph-H), 7.67 (td, J = 7.7,
2.6 Hz, 1 H, Ph-H), 7.31-7.39 (m, 5 H, Ph-H), 6.33 (d, J = 2.6 Hz 1 H, H-10),
6.25 (brt, 1 H, H-13), 5.63 (d, J = 6.2 Hz, 1 H, H-2), 5.33 (dd like, J = 7.7
Hz, 2 H, CONH), 4.74 (broad s, 1 H, H-5), 4.62 (s, 1 H, H-2′), 4.23 (d, J = 7.7 Hz, 1
H, H-20a), 4.12 (d, J = 6.2 Hz, 1 H, H-3), 4.01 (d, J = 7.7 Hz, 1 H, H-20b),
2.46 (dt like, J = 15.7, 1 H, H-6a), 2.37 (s, 3 H, CH 3CO in
C-4), 2.24-2.33 (m, 3 H, H-19a and H-14), 2.21 (s, 3 H, CH 3CO
in C-10), 2.11 (d, J = 16.1 Hz, 1 H, H-6b), 1.85 (s, 3 H, CH 3
in C-18), 1.66 (t like, 1 H, H-19b), 1.36 (m, 1 H, H-7), 1.25-1.28 (m, 15 H, Me 3C,
H-16 and H-17)。
去苯甲酰基 -2-( 间 - 叠氮基苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -1h): 1H NMR (400 MHz, CDCl3)
δ 7.93 (d, J = 7.8 Hz, 1 H,
Ph-H), 7.86 (s, 1 H, Ph-H), 7.48 (t, J = 7.8 Hz, 1 H, Ph-H), 7.22-7.41 (m, 6 H,
Ph-H), 6.33 (s, 1 H, H-10), 6.26 (brt, 1 H, H-13), 5.66 (d, J = 7.8 Hz, 1 H,
H-2), 5.34 (d like, 1 H, CONH), 5.32 (broad d, 1 H, H-3′), 4.74 (d, 1 H, J = 3.4
Hz, H-5), 4.60 (s, 1 H, H-2′), 4.33 (d, J = 8.8 Hz, 1 H, H-20a), 4.10 (d, J = 7.3 Hz, 1 H, H-3), 4.03
(d, J = 8.8 Hz, 1 H, H-20b), 3.25 (broad s, 1H), 2.46 (dt like, J = 16.1, 1 H,
H-6a), 2.39 (s, 3 H, CH 3CO- in C-4), 2.23-2.26 (m, 3 H, H-19a
and H-14), 2.21 (s, 3 H, CH 3CO- in C-10), 2.11 (d, J = 16.1
Hz, 1 H, H-6b), 1.85 (s, 3 H, -CH 3 in C-18), 1.65 (t like, 1
H, H-19b), 1.38 (m, 1 H, H-7), 1.25-1.26 (m, 15 H, Me 3C-,
H-16 and H-17)。
侧链通式(Ⅵ) 所示化合物与母环通式(Ⅴ)所示化合物对接产物的制备方法与(Ⅰ -1a~ Ⅰ -1h)相似:
3'-(2, 2- 二氟乙烯基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -2a). 1H NMR (400 MHz, CDCl3)
δ 8.17 (d, J = 7.4 Hz, 2 H,
Ph-H), 7.61 (t, J = 7.4 Hz, 1 H, Ph-H), 7.51 (t, J = 7.8 Hz, 2 H, Ph-H), 6.33
(s, 1 H, H-10), 6.27 (broad t, 1 H, H-13), 5.66 (d, J = 7.8 Hz, 1 H, H-2), 4.90
(d, J = 2.7 Hz, 1 H), 4.74 (d, J = 3.5 Hz, 1 H, H-5), 4.62-4.56 (m, 1 H, H-2′), 4.32 (d, J = 8.6 Hz, 1
H, H-20a), 4.28 (broad s, 1 H), 4.09 (d, J = 7.8 Hz, 1 H, H-3), 4.04 (d, J =
8.2 Hz, 1 H), 3.44 (broad s, 1 H, -OH), 2.50-2.42 (m, 2 H, H-6a and
H-14a), 2.39 (s, 3 H, CH 3CO- in C-4), 2.27-2.21 (m, 2 H, H-19a
and H-14b), 2.21 (s, 3 H, CH 3CO- in C-10), 2.11 (d, J = 16.0
Hz, 1 H, H-6b), 1.86 (s, 3 H, -CH 3 in C-18), 1.67 (t like, 1
H, H-19b), 1.37 (m, 1 H, H-7), 1.25 (m, 15 H, Me 3C-, H-16 and
H-17)。
甲基 -1- 丙烯基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -2b). 1H NMR (400 MHz, CDCl3)
δ 8.15 (d, J = 7.3 Hz, 2 H,
Ph-H), 7.59 (t, J = 7.3 Hz, 1 H, Ph-H), 7.47 (t, J = 7.7 Hz, 2 H, Ph-H), 6.33
(s, 1 H, H-10), 6.19 (broad t, J = 8.8 Hz, 1 H, H-13), 5.65 (d, J = 7.7 Hz, 1
H), 4.73 (d, J = 3.7 Hz, 1 H), 4.59 (d, J = 9.9 Hz, 1 H), 4.29 (d, J = 8.4 Hz,
1 H), 4.16 (d, J = 4.4 Hz, 2 H), 4.08 (t like, J = 8.8, 7.7 Hz, 2 H), 3.26 (d,
J = 5.1 Hz, 1 H), 2.44-2.49 (m, 2 H), 2.35 (s, 3 H, CH 3CO- in
C-4), 2.32-2.38 (m, 1 H), 2.21-2.25 (m, 2 H), 2.20 (s, 3 H, CH 3CO-
in C-10), 2.09 (d, J = 16.1 Hz, 1 H, H-6b), 1.86 (s, 3 H, CH 3
in C-18), 1.68 (m, 6 H), 1.33-1.38 (m, 1 H, H-7), 1.24-1.28 (m, 15 H, Me 3C-,
H-16 and H-17), 0.99 (s, 3 H), 0.98 (s, 3 H)。
甲基丙基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -2c). 1H NMR (400 MHz, CDCl3)
δ 8.14 (d, J = 7.5 Hz, 2 H,
Ph-H), 7.60 (t, J = 7.5 Hz, 1 H, Ph-H), 7.49 (t, J = 7.5 Hz, 2 H, Ph-H), 6.33
(s, 1 H, H-10), 6.16 (broad t, 1 H, H-13), 5.66 (d, J = 7.5 Hz, 1 H), 5.32 (d,
J = 7.5 Hz, 1 H ), 4.73-4.79 (m, 2 H), 4.30 (d, J = 8.7 Hz, 1 H), 4.19 (broad
s, 1 H), 4.02-4.14 (m, 2 H), 3.38 (broad s, 1 H), 2.36-2.48 (m, 3 H), 2.34 (s,
3 H), 2.21-2.24 (m, 1 H), 2.20 (s, 3 H), 2.09 (d, J = 17.4 Hz, 1 H), 1.87 (s, 3
H), 1.77 (s, 6 H), 1.66 (m, 1 H), 1.32 (m, 9 H), 1.26 (s, 3 H), 1.23 (s, 3 H)。
去叔丁氧羰基-3'-(2- 甲基丙烯基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅱ -a) :
将已制备的3'-(2-甲基丙烯基)-7-脱氧-7β, 8β-亚甲基-10-乙酰氧基-多西紫杉醇(Ⅰ -2b) (1.8 g, 2.22 mmol)溶于CH2Cl2
(30 mL),0 °C 下加入三氟乙酸的二氯甲烷溶液 (10 mL, 25% total volume),继续反应1.5 h,TLC检测反应物原料剩余后, 用饱和碳酸氢钠终止反应,水相用CH2Cl2
(100 mL × 3)萃取,合并有机相,依次用饱和碳酸氢钠水溶液(100 mL × 3)、饱和氯化钠水溶液(100 mL × 3)洗涤,无水硫酸钠干燥,浓缩,粗产物经柱层析分离(石油醚/乙酸乙酯 = 1/3)得到白色鱼鳞状固体3'-N-去叔丁氧羰基-3'-(2-甲基丙烯基)-7-脱氧-7β, 8β-亚甲基-10-乙酰氧基-多西紫杉醇(Ⅱ -a) (1.48 g, 94% yield):
1H NMR (400 MHz, CDCl3) δ 8.11 (d, J = 7.3 Hz, 2 H, Ph-H),
7.62 (t, J = 7.3 Hz, 1 H, Ph-H), 7.48 (t, J = 7.8 Hz, 2 H, Ph-H), 6.34 (s, 1 H,
H-10), 6.19 (broad t, 1 H, H-13), 5.67 (d, J = 7.8 Hz, 1 H), 5.25 (d, J = 8.8
Hz, 1 H ), 4.74 (d, J = 8.8 Hz, 1 H), 4.32 (d, J = 8.8 Hz, 1 H), 4.05-3.89 (m,
3 H), 2.47 (dt, J = 16.1, 4.4 Hz, 1 H), 2.34 (s, 3 H), 2.24-2.26 (m, 2 H), 2.21
(s, 3 H), 2.11 (d, J = 16.1 Hz, 1 H), 1.89 (s, 3 H), 1.73 (s, 3 H), 1.71 (s, 3
H), 1.66 (t like, 1 H), 1.38 (m, 1 H), 1.28 (s, 3 H), 1.25 (s, 3 H)。
根据相同的方法可以制备C-3'氨基脱Boc保护基的中间体(Ⅱ -b):
3'-N- 去叔丁氧羰基-3'-(2- 甲基丙基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅱ -b): 1H NMR (400 MHz, CDCl3)
δ 8.16 (d, J = 7.8 Hz, 2 H,
Ph-H), 7.68 (t, J = 7.3 Hz, 1 H, Ph-H), 7.54 (t, J = 7.3 Hz, 2 H, Ph-H), 6.39
(s, 1 H, H-10), 6.28 (broad t, J = 8.8 Hz, 1 H, H-13), 5.72 (d, J = 7.3 Hz, 1
H, H-2), 4.79 (d like, 1 H), 4.37 (d, J = 8.8 Hz, 1 H), 4.16 (d, J = 7.3 Hz, 1
H), 4.09 (d, J = 7.8 Hz, 2 H), 4.08 (t like, J = 8.8, 7.7 Hz, 2 H), 3.30 (broad
s, 1 H), 2.56 (dt like, J = 7.8 Hz, 1 H, H-6a), 2.39 (s, 3 H, CH 3CO
in C-4), 2.29-2.41 (m, 5 H), 2.26 (s, 3 H, CH 3CO in C-10), 2.16
(d, J = 16.1 Hz, 1 H, H-6b), 1.96 (s, 3 H, CH 3 in C-18), 1.71
(t like, 1 H), 1.38-1.43 (m, 1 H, H-7), 1.32 (s, 3 H), 1.30 (s, 3 H), 1.05 (d,
J = 7.3 Hz, 3 H), 1.01 (d, J = 7.3 Hz, 3 H)。
去叔丁氧羰基-3'-N-((E)-2- 甲基 -2- 丁烯酰基 )-(2- 甲基 -1- 丙烯基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -3a):
将制备得到的3'-N-去叔丁氧羰基-3'-(2-甲基丙烯基)-7-脱氧-7β, 8β-亚甲基-10-乙酰氧基-多西紫杉醇(Ⅱ -a) (78.0 mg, 0.11 mmol)、三乙胺 (30.7 µL, 0.22 mmol)溶于CH2Cl2
(10 mL),加入缩合剂(EDC·HCl) (31.8 mg, 0.17 mmol) 和 trans-2,
3-dimethylacrylic acid (14.3 mg, 0.17 mmol),室温搅拌4 h,TLC检测反应物原料剩余后, 用饱和氯化铵终止反应,水相用CH2Cl2 (30 mL × 3)萃取,合并有机相,依次用饱和碳酸氢钠水溶液(20 mL × 3)、饱和氯化钠水溶液(20 mL × 3)洗涤,无水硫酸钠干燥,浓缩,粗产物经柱层析分离(石油醚/乙酸乙酯 = 3/1)得到白色泡沫状固3'-N-去叔丁氧羰基-3'-N-((E)-2-甲基-2-丁烯酰基)-(2-甲基-1-丙烯基)-7-脱氧-7β, 8β-亚甲基-10-乙酰氧基-多西紫杉醇(Ⅰ -3a) (74.9 mg, 86% yield):
1H NMR (400 MHz, CDCl3) δ 8.14 (d, J = 7.8 Hz, 2 H, Ph-H),
7.47 (t, J = 7.0 Hz, 1 H, Ph-H), 7.47 (t, J = 7.8 Hz, 2 H, Ph-H), 6.36 (dd, J =
14.1, 7.0 Hz, 1 H), 6.32 (s, 1 H, H-10), 6.15 (broad t, J = 8.6 Hz, 1 H, H-13),
5.99 (d, J = 8.2 Hz, 1 H), 5.39 (d, J = 9.0 Hz, 1 H, -CONH-), 5.09 (td,
J = 8.2, 3.1 Hz, 1 H), 4.73 (d, J = 3.5 Hz, 1 H, H-5), 4.30 (d, J = 8.6 Hz, 1 H,
H-20a), 4.27 (s, 1 H), 4.09-4.07 (m, 2 H, H-20b and H-3), 3.76 (broad s, 1H),
2.49-2.33 (m, 3 H), 2.35 (s, 3 H, CH 3CO-), 2.26-2.22 (m, 1
H), 2.20 (s, 3 H, CH 3CO-), 2.09 (d, J = 15.7 Hz, 1 H, H-6b),
1.85 (s, 3 H-CH 3 in C-18), 1.81-1.68 (m, 13 H), 1.39-1.31 (m,
1 H, H-7), 1.29-1.21 (m, 6 H, -CH3 in C-16 and C-17)。
根据相同的方法可以制备其他的C-3'氨基N取代的化合物(Ⅰ -3b~ Ⅰ -3c):
3'-N- 去叔丁氧羰基-3'-N-((E)-2- 丁烯酰基 )-(2- 甲基 -1- 丙烯基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -3c): 1H NMR (400 MHz, CDCl3)
δ 8.14 (d, J = 7.0 Hz, 2 H,
Ph-H), 7.60 (t, J = 7.4 Hz, 1 H, Ph-H), 7.47 (t, J = 7.8 Hz, 2 H, Ph-H),
6.75-6.69 (m, 1H), 6.32 (s, 1 H, H-10), 6.15 (broad t, 1 H, H-13), 5.76 (d, J =
8.6 Hz, 1 H), 5.71 (s, 1H), 5.67 (d, J = 7.8 Hz, 1 H), 5.39 (d, J = 9.0 Hz, 1
H, -CONH-), 5.09 (td, J = 8.6, 2.7 Hz, 1 H), 4.73 (d, J = 3.5 Hz, 1 H,
H-5), 4.30 (d, J = 8.6 Hz, 1 H, H-20a), 4.26 (d, J = 2.3 Hz, 1 H), 4.08 (d, J =
8.2 Hz, 2 H, H-20b and H-3), 2.49-2.37 (m, 3 H), 2.34 (s, 3 H CH 3CO-),
2.25-2.21 (m, 1 H), 2.20 (s, 3 H CH 3CO-), 2.09 (d, J = 14.9
Hz, 1 H, H-6b), 1.85 (s, 3 H, -CH3 in C-18), 1.77 (s, 9 H),
1.68-1.65 (m, 1 H, H-19b), 1.36 (m, 1 H, H-7), 1.26-1.23 (m, 6 H, -CH3
in C-16 and C-17)。
去叔丁氧羰基-3'-N-((E)-2- 甲基 -2- 丁烯酰基 )-(2- 甲基丙基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -3d): 1H NMR (400 MHz, CDCl3)
δ 8.16 (d, J = 7.8 Hz, 2 H,
Ph-H), 7.59 (t, J = 7.4 Hz, 1 H, Ph-H), 7.47 (t, J = 7.8 Hz, 2 H, Ph-H), 6.32
(s, 1 H, H-10), 6.29 (dd, J = 13.7, 7.0 Hz, 1 H), 6.16 (broad t, 1 H, H-13),
5.79 (d, J = 9.4 Hz, 1 H), 5.67 (d, J = 7.8 Hz, 1 H, H-2), 4.73 (d, J = 3.1 Hz,
1 H, H-5), 4.49 (m, 1 H), 4.29 (d, J = 8.6 Hz, 1 H, H-20a), 4.23 (broad s, 1 H,
H-2'), 4.11-4.07 (m, 2 H, H-20b and H-3), 3.59 (broad s, 1 H, -OH),
2.50-2.33 (m, 3 H), 2.39 (s, 3 H, CH 3CO-), 2.28-2.20 (m, 1
H), 2.20 (s, 3 H, CH 3CO-), 2.10 (d, J = 15.7 Hz, 1 H, H-6b),
1.85 (s, 3 H, -CH 3 in C-18), 1.85-1.67 (m, 8 H), 1.43-1.37
(m, 3 H), 1.27-1.23 (m, 6 H, -CH3 in C-16 and C-17), 1.00 (s,
3 H), 0.98 (s, 3 H)。
去叔丁氧羰基-3'-N-((E)-2- 丁烯酰基 )-(2- 甲基丙基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -3e): 1H NMR (400 MHz, CDCl3)
δ 8.16 (d, J = 7.0 Hz, 2 H,
Ph-H), 7.59 (t, J = 7.4 Hz, 1 H, Ph-H), 7.47 (t, J = 7.8 Hz, 2 H, Ph-H),
6.68-6.63 (m, 1 H, CH3CH=CH-), 6.32 (s, 1 H, H-10), 6.16
(broad t, J = 8.6 Hz, 1 H, H-13), 5.72 (dd, J = 15.3, 1.6 Hz, 1 H), 5.67 (d, J
= 7.4 Hz, 1 H, H-2), 5.58 (d, J = 9.0 Hz, 1 H, -CONH-), 4.73 (d, J = 3.6
Hz, 1 H, H-5), 4.48 (m, 1 H), 4.29 (d, J = 8.6 Hz, 1 H, H-20a), 4.22 (broad s,
1 H, H-2'), 4.11 (d, J = 8.6 Hz, 1 H, H-20b), 4.07 (d, J = 7.8 Hz, 1 H, H-3),
3.61 (broad s, 1 H, -OH), 2.50-2.44 (m, 3 H), 2.37 (s, 3 H, CH 3CO-),
2.26-2.20 (m, 1 H), 2.20 (s, 3 H, CH 3CO-), 2.10 (d, J = 15.7
Hz, 1 H, H-6b), 1.84 (s, 3 H, -CH 3 in C-18), 1.84-1.59 (m, 5
H), 1.40-1.35 (m, 3 H), 1.27-1.23 (m, 6 H, -CH3 in C-16 and
C-17), 1.00-0.98 (m, 6 H)。
去叔丁氧羰基-3'-N-(3, 3'- 二甲基丙烯酰基 )-(2- 甲基丙基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -3f): 1H NMR (400 MHz, CDCl3)
δ 8.15 (d, J = 7.4 Hz, 2 H,
Ph-H), 7.58 (t, J = 7.4 Hz, 1 H, Ph-H), 7.46 (t, J = 7.8 Hz, 2 H, Ph-H), 6.32
(s, 1 H, H-10), 6.16 (broad t, 1 H, H-13), 5.66 (d, J = 7.4 Hz, 1 H, H-2), 5.50
(s, 1 H), 5.45 (d, J = 9.4 Hz, 1 H, -CONH-), 4.73 (d, J = 3.5 Hz, 1 H,
H-5), 4.46 (m, 1 H), 4.29 (d, J = 8.6 Hz, 1 H, H-20a), 4.21 (d, J = 1.6 Hz, 1
H), 4.10-4.07 (m, 2 H, H-20b and H-3), 2.51-2.34 (m, 3 H), 2.38 (s, 3 H, CH 3CO-),
2.56-2.20 (m, 2 H), 2.20 (s, 3 H, CH 3CO-), 2.10 (d, J = 15.7
Hz, 1 H, H-6b), 1.91 (s, 3 H), 1.85 (s, 3 H, -CH 3 in C-18),
1.77 (s, 3 H), 1.77-1.65 (m, 2 H), 1.41-1.34 (m, 3 H), 1.25 (m, 6 H, -CH3
in C-16 and C-17), 1.00-0.97 (m, 6 H)。
甲氧基
-7-
脱氧
-7
β
, 8
β
-
亚甲基
-10-
乙酰氧基
-
多西紫杉醇
(
Ⅰ
-4a):
1H NMR (400 MHz, CDCl3)δ 8.14 (d, J = 7.4 Hz, 2 H,
Ph-H), 7.60 (t, J = 7.4 Hz, 1 H, Ph-H), 7.50 (t, J = 7.8 Hz, 2 H, Ph-H),
7.42-7.29 (m, 5 H, Ph-H), 6.31 (brt, J = 8.6 Hz, 1 H, H-13), 5.66 (d, J = 7.4
Hz, 1 H, H-2), 5.37 (d, J = 9.4 Hz, 1 H, CONH), 5.30 (brd, 1 H, H-3′), 4.95 (s, 1 H, H-10),
4.72 (d, J = 3.5 Hz, 1 H, H-5), 4.60 (s, 1 H, H-2′), 4.30 (d, J = 9.0 Hz, 1 H, H-20a),
4.09 (d, J = 7.4 Hz, 1 H, H-3), 4.04 (d, J = 8.6 Hz, 1 H, H-20b), 3.45 (s, 3 H,
-OCH3 ), 3.33 (brs, 1 H, -OH), 2.43-2.29 (m, 3 H, H-6a and
H-14), 2.37 (s, 3 H, CH 3CO in C-4), 2.26-2.18 (m, 1 H,
H-19a), 2.21 (s, 3 H, CH 3CO in C-10), 2.10 (d, J = 16.0 Hz, 1
H, H-6b), 1.86 (s, 3 H, -CH 3 in C-18), 1.63 (t like, 1 H,
H-19b), 1.36-1.30 (m, 1 H, H-7), 1.29 (s, 9 H, Me 3C-), 1.27
(s, 3 H, -CH3 ), 1.23 (s, 3 H, -CH3 );
7-
脱氧
-7
β
, 8
β
-
亚甲基
-10-
乙酰氧基
-
多西紫杉醇
(
Ⅰ
-4b):
1H NMR (400 MHz, CDCl3)δ 8.15 (d, J = 7.4 Hz, 2 H,
Ph-H), 7.60 (t, J = 7.4 Hz, 1 H, Ph-H), 7.51 (t, J = 7.8 Hz, 2 H, Ph-H),
7.42-7.31 (m, 5 H, Ph-H), 6.29 (brt, 1 H, H-13), 5.66 (d, J = 7.8 Hz, 1 H,
H-2), 5.41 (d, J = 9.4 Hz, 1 H, CONH), 5.29 (brd, 1 H, H-3′), 5.01 (s, 1 H, H-10),
4.73 (d, J = 3.1 Hz, 1 H, H-5), 4.61 (s, 1 H, H-2′), 4.32 (d, J = 8.6 Hz, 1 H, H-20a),
4.24 (s, 1 H), 4.13 (d, J = 7.4 Hz, 1 H, H-3), 4.06 (d, J = 8.6 Hz, 1 H,
H-20b), 3.33 (brs, 1 H, -OH), 2.44-2.31 (m, 3 H, H-6a and H-14), 2.38 (s, 3 H,
CH 3CO in C-4), 2.24-2.20 (m, 1 H, H-19a), 2.21 (s, 3 H, CH 3CO
in C-10), 2.13 (d, J = 16.0 Hz, 1 H, H-6b), 1.85 (s, 3 H, -CH 3
in C-18), 1.77 (t like, 1 H, H-19b), 1.44 (m, 1 H, H-7), 1.28 (s, 9 H, Me 3C-),
1.26 (s, 3 H, -CH3 ), 1.20 (s, 3 H, -CH3 )。
二氟乙烯基
)-2-
去苯甲酰基
-2-(
间
-
叠氮基苯甲酰基
)-7-
脱氧
-7
β
, 8
β
-
亚甲基
-10-
乙酰氧基
-
多西紫杉醇
(
Ⅰ
-5a):
1H NMR (400 MHz, CDCl3)δ 7.93 (d, J = 7.6 Hz, 1 H,
Ph-H), 7.87 (s, J = 7.4 Hz, 1 H, Ph-H), 7.49 (t, J = 8.0 Hz, 1 H, Ph-H), 7.23
(dd, J = 8.0, 1.6 Hz, 1 H, Ph-H), 6.32 (s, 1H, H-10), 6.25 (brt, 1 H, H-13),
5.66 (d, J = 7.6 Hz, 1 H, H-2), 4.90 (m, 2 H, CONH and H-3′), 4.75 (d, J = 3.6 Hz, 1
H, H-5), 4.56 (dd, J = 24.8, 8.0 Hz, 1 H, CF2CH), 4.37-4.33
(m, 2 H, H-20a and H-2′), 4.26 (s, 1 H), 4.10 (d, J = 7.6 Hz, 1 H, H-3), 4.03 (d, J = 8.4 Hz, 1
H, H-20b), 3.45 (brs, 1 H), 2.88 (s, 1 H), 2.50-2.35 (m, 3 H, H-6a and H-14),
2.39 (s, 3 H, CH 3CO in C-4), 2.29-2.23 (m, 1 H, H-19a), 2.20
(s, 3 H, CH 3CO in C-10), 2.11 (d, J = 16.4 Hz, 1 H, H-6b),
1.86 (s, 3 H, -CH 3 in C-18), 1.65 (t like, 1 H, H-19b),
1.40-1.35 (m, 1 H, H-7), 1.26 (s, 9 H, Me 3C-), 1.24 (s, 6 H,
-CH3 )。
去叔丁氧羰基-3'-N-((E)-2- 丁烯酰基 )-3'-(2, 2- 二氟乙烯基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -5b): 1H NMR (400 MHz, CDCl3)δ 8.18 (d, J = 7.2 Hz, 2 H,
Ph-H), 7.60 (t, J = 7.2 Hz, 1 H, Ph-H), 7.52 (t, J = 7.2 Hz, 2 H, Ph-H),
6.72-6.63 (m, 1 H, CH3CHCH), 6.32 (s, 1 H, H-10), 6.24 (brt,
1 H, H-13), 5.84 (d, J = 8.8 Hz, 1 H, CH3CHCH), 5.71-5.65 (m,
2 H), 5.21 (brt, J = 9.4 Hz, 1 H, CONH), 4.73 (d, J = 4.0 Hz, 1 H, H-5),
4.66 (ddd, J = 24.8, 10.0, 1.6 Hz, 1 H), 4.34 (d, J = 2.4 Hz, 1 H), 4.31 (d, J
= 8.8 Hz, 1 H, H-20a), 4.08 (d, J = 6.8 Hz, 1 H, H-3), 4.07 (d, J = 8.8 Hz, 1
H, H-20b), 2.87 (s, 1 H), 2.50-2.39 (m, 3 H, H-6a and H-14), 2.41 (s, 3 H, CH 3CO
in C-4), 2.25-2.18 (m, 1 H, H-19a), 2.20 (s, 3 H, CH 3CO in
C-10), 2.10 (d, J = 14.8 Hz, 1 H, H-6b), 1.85 (s, 3 H, -CH 3
in C-18), 1.73 (dd, J = 7.2, 1.6 Hz, 3 H, CH3 CHCH),
1.68 (t like, 1 H, H-19b), 1.41-1.35 (m, 1 H, H-7), 1.26-1.23 (m, 6 H, -CH3 )。
去叔丁氧羰基-3'-N-((E)-2- 丁烯酰基 )-3'-(2, 2- 二氟乙烯基 )-2- 去苯甲酰基 -2-( 间 - 叠氮基苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -5c):
1H NMR (400 MHz, CDCl3)δ 7.93 (d, J = 7.6 Hz, 1 H,
Ph-H), 7.87 (s, J = 7.4 Hz, 1 H, Ph-H), 7.50 (t, J = 8.0 Hz, 1 H, Ph-H), 7.23
(dd, J = 8.0, 1.6 Hz, 1 H, Ph-H),, 6.68-6.58 (m, 1 H, CH3CHCH),
6.32 (s, 1 H, H-10), 6.21 (brt, 1 H, H-13), 5.86 (d, J = 8.8 Hz, 1 H, CH3CHCH),
5.71 (d, J = 1.6 Hz, 1 H), 5.66 (d, J = 8.0 Hz, 2 H), 5.19 (brt, J = 9.2 Hz, 1
H, CONH), 4.75 (d, J = 4.0 Hz, 1 H, H-5), 4.65 (ddd, J = 24.4, 10.0, 1.6
Hz, 1 H), 4.37-4.32 (m, 3 H), 4.10-4.04 (m, H-3 and H-20b), 4.07 (d, J = 8.8
Hz, 1 H, H-20b), 2.88 (s, 1 H), 2.52-2.39 (m, 3 H, H-6a and H-14), 2.41 (s, 3
H, CH 3CO in C-4), 2.30-2.22 (m, 1 H, H-19a), 2.20 (s, 3 H, CH 3CO
in C-10), 2.11 (d, J = 16.4 Hz, 1 H, H-6b), 1.85 (s, 3 H, -CH 3
in C-18), 1.74 (dd, J = 6.8, 1.6 Hz, 3 H, CH3 CHCH),
1.67 (dd, J = 7.2, 5.6 Hz, 1 H, H-19b), 1.45-1.30 (m, 1 H, H-7), 1.26-1.23 (m,
6 H, -CH3 )。
去叔丁氧羰基-3'-N-((E)-4, 4, 4- 三氟 -2- 丁烯酰基 )-3'-(2, 2- 二氟乙烯基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -5d): 1H NMR (400 MHz, CDCl3)δ 8.20 (d, J = 7.6 Hz, 2 H,
Ph-H), 7.60 (t, J = 7.2 Hz, 1 H, Ph-H), 7.52 (t, J = 7.6 Hz, 2 H, Ph-H),
6.51-6.22 (m, 3 H, CF3CHCH, CF3CHCH and
H-13), 6.32 (s, 1 H, H-10), 5.65 (d, J = 7.6 Hz, 1 H), 5.31 (brt, J = 9.4 Hz, 1
H, CONH), 4.73 (d, J = 3.6 Hz, 1 H, H-5), 4.64 (dd, J = 24.4, 10.0, 1
H), 4.35-4.31 (m, 3 H), 4.10-4.06 (m, 2 H), 3.56 (s, 1 H), 2.88 (s, 1 H),
2.52-2.43 (m, 3 H, H-6a and H-14), 2.41 (s, 3 H, CH 3CO in
C-4), 2.29-2.09 (m, 2 H), 2.20 (s, 3 H, CH 3CO in C-10), 1.86
(s, 3 H, -CH 3 in C-18), 1.68 (t like, 1 H, H-19b), 1.47-1.37
(m, 1 H, H-7), 1.27-1.23 (m, 6 H, -CH3 )。
去叔丁氧羰基-3'-N-((E)-4, 4, 4- 三氟 -2- 丁烯酰基 )-3'-(2, 2- 二氟乙烯基 )-2- 去苯甲酰基 -2-( 间 - 叠氮基苯甲酰基 )-7- 脱氧 -7 β , 8 β - 亚甲基 -10- 乙酰氧基 - 多西紫杉醇 ( Ⅰ -5e): 1H NMR (400 MHz, CDCl3)
δ 7.96 (d, J = 7.9 Hz, 1 H,
Ph-H), 7.90 (d, J = 1.6 Hz, 1 H, Ph-H), 7.50 (t, J = 7.9 Hz, 1 H, Ph-H), 7.23
(dd, J = 7.9, 1.6 Hz, 1 H, Ph-H), 6.44-6.38 (m, 2 H), 6.32 (s, 1 H, H-10), 6.23
(brt, 1 H, H-13), 5.65 (d, J = 7.9 Hz, 1 H), 5.31 (brt, J = 9.5 Hz, 1 H, CONH),
4.75 (d, J = 4.0 Hz, 1 H, H-5), 4.64 (dd, J = 24.4, 10.0, 1 H), 4.37-4.34 (m, 3
H), 4.09-4.04 (m, 2 H), 2.89 (s, 1 H), 2.50-2.31 (m, 3 H, H-6a and H-14), 2.42
(s, 3 H, CH 3CO in C-4), 2.24-2.09 (m, 2 H), 2.20 (s, 3 H, CH 3CO
in C-10), 1.85 (s, 3 H, -CH 3 in C-18), 1.66 (t like, 1 H,
H-19b), 1.41-1.30 (m, 1 H, H-7), 1.27-1.23 (m, 6 H, -CH3 )。
最终合成的部分紫杉烷衍生物对KB(人口腔鳞癌细胞株)、KB/VCR(耐长春新碱的人口腔鳞癌耐药细胞株)、MCF-7(人乳腺癌细胞株)、MCF-7/ADR(耐阿霉素的人乳腺癌耐药细胞株)进行了体外抗肿瘤活性评价实验,活性测试方法如下:
1.实验材料:KB、KB/VCR、MCF-7、MCF-7/ADR四种细胞株均为中科院上海药物所药理实验室提供;丙酮酸钠、谷氨酰胺、磺酰罗丹明B (sulforhodamin B,SRB)、二甲基亚砜(DMSO) (均为美国Sigma公司产品);小牛血清(FBS)、DMEM培养基、MEM培养基干粉和胰蛋白酶(均为美国GIBCO公司产品);三氯乙酸、醋酸等其他试剂均为分析纯(国药集团化学试剂有限公司);水为注射用生理盐水。
2.细胞培养:KB和KB/VCR细胞培养培养液组分为:MEM+谷氨酰胺(1%)+丙酮酸钠(1%)+小牛血清(10%);MCF-7细胞培养培养液组分为:DMEM+小牛血清(10%);MCF-7/ADR细胞培养培养液组分为:MEM+胰岛素(1%)+丙酮酸钠(1%)+小牛血清(10%)。所有细胞培养于5% CO2,37℃饱和湿度的培养箱中;
3.SRB法:取对数生长期KB、KB/VCR、MCF-7、MCF-7/ADR四种细胞株,用0.125% 胰酶消化、吹匀成单细胞悬液,计数活细胞。KB、KB/VCR、MCF-7、MCF-7/ADR细胞分别以6~7×103、4~5×103、4~5×103、7~8×103个细胞/孔接种于96孔板中(每孔100 µL)。细胞培养24 h后,实验组每孔加入事先用生理盐水配好的8个梯度浓度的化合物,每种药相同浓度下平行测三组。培养72 h后,每孔加入4℃预冷的10%的三氯醋酸(TCA, 100 µL),在4℃放置1 h固定细胞。接着将培养板各孔用去离子水洗涤6~8遍,于电热鼓风干燥箱内37℃干燥至无湿痕。然后每孔加0.4%的SRB(以1%的醋酸配制)100 µL,室温下避光染色15 min。弃去各孔内液体后,用1%的醋酸洗涤6~8遍,然后于电热鼓风干燥箱内37℃下干燥至无湿痕后,每孔加入10 mmol/L的 Tris (150 µL/孔)溶解SRB,用酶标仪以515 nm的波长测定OD值及计算半数抑制浓度IC50值。细胞增殖抑制率按下式计算:细胞增殖抑制率(%) = (1-实验组OD值)/对照组OD值)×100,将每种化合物三组计算所得抑制率取平均值。重复上述操作一次,将两次所得半数抑制浓度IC50值取平均值,测试结果总结如下表所示:
|
KB (nM) |
KB/VCR (nM) |
R/S |
MCF-7 (nM) |
MCF-7/ADR (nM) |
R/S |
阿霉素 |
NT |
NT |
— |
32.4 |
7.6 (µM) |
233.4 |
长春新碱 |
11.1 |
588.0 |
53.0 |
NT |
NT |
— |
紫杉醇 |
10.3 |
4.0 (µM) |
388.3 |
9.6 |
>100.0 (µM) |
>10000 |
多烯紫杉醇 |
7.3 |
169.7 |
23.2 |
1.7 |
1530.5 |
900.3 |
Larotaxel |
15.8 |
57.5 |
3.6 |
18.6 |
280.8 |
15.1 |
Ⅰ-1a
|
30.4 |
167.1 |
5.5 |
71.0 |
667.9 |
9.4 |
Ⅰ-1b
|
13.7 |
63.0 |
4.7 |
37.4 |
223.2 |
6.0 |
Ⅰ-1c
|
37.2 |
237.6 |
6.4 |
91.8 |
670.3 |
7.3 |
Ⅰ-1d
|
17.5 |
93.0 |
5.3 |
35.1 |
231.5 |
6.6 |
Ⅰ-1e
|
20.9 |
81.0 |
3.9 |
26.7 |
239.8 |
9.0 |
Ⅰ-1f
|
18.2 |
50.3 |
2.8 |
28.7 |
175.6 |
6.1 |
Ⅰ-1g
|
25.7 |
125.6 |
4.9 |
20.5 |
449.6 |
21.9 |
Ⅰ-1h
|
11.8
|
42.0
|
3.6
|
23.8
|
56.9
|
2.4
|
Ⅰ-2b
|
2.5
|
14.2
|
5.7
|
1.6
|
10.6
|
6.6
|
Ⅰ-3a
|
13.6
|
41.4
|
3.0
|
6.1
|
48.1
|
7.9
|
Ⅰ-3c
|
3.2
|
19.4
|
6.1
|
2.4
|
14.8
|
6.2
|
Ⅰ-3b
|
0.038
|
10.7
|
281.6
|
0.039
|
9.2
|
236.7
|
Ⅰ-2c
|
59.2 |
103.3 |
1.7 |
79.7 |
169.3 |
2.1 |
Ⅰ-3d
|
15.4 |
95.6 |
6.2 |
13.5 |
308.1 |
22.8 |
Ⅰ-3f
|
0.1
|
12.2
|
122.0
|
0.24
|
45.5
|
189.6
|
Ⅰ-3e |
0.6 |
23.3 |
38.3 |
1.7 |
138.7 |
81.6 |
Ⅰ-2a
|
3.0
|
8.1
|
2.7
|
2.8
|
25.6
|
9.1
|
Ⅰ-4a
|
3.2 |
65.9 |
20.6 |
9.0 |
269.9 |
30.0 |
Ⅰ-4b
|
2.4 |
22.5 |
9.4 |
7.2 |
165.8 |
12.3 |
Ⅰ-5a
|
6.1
|
22.9
|
3.8
|
35.9
|
56.6
|
1.6
|
Ⅰ-5b
|
2.5 |
25.8 |
10.3 |
1.4 |
417.6 |
298.3 |
Ⅰ-5c
|
5.3
|
15.2
|
2.9
|
6.0
|
37.1
|
6.2
|
Ⅰ-5d
|
4.7 |
298.0 |
64.3 |
22.7 |
4.7 (µM) |
205.2 |
Ⅰ-5e
|
10.5 |
95.4 |
9.1 |
24.6 |
394.4 |
16.0 |
从生物活性测试结果可以看出,化合物该类化合物多数能有效地抑制肿瘤细胞的生长,尤其是针对多药耐药(MDR)肿瘤细胞(MCF-7/ADR、KB/VCR),所示的9个化合物Ⅰ -1h、Ⅰ -2a、Ⅰ -2b、Ⅰ -3a、Ⅰ -3b、Ⅰ -3c、Ⅰ -3f、Ⅰ -5a、Ⅰ -5c与紫杉醇(Paclitaxel, Taxol®)相比具有3~5个数量级活性的提高,与多西紫杉醇(Docetaxel, Taxotere®)相比具有1~3个数量级活性的提高,与拉洛他赛(Larotaxel, XRP9881)相比具有1~2个数量级活性的提高,因而,该紫杉烷类衍生物可用于临床上对高表达P-糖蛋白的多药耐药肿瘤的治疗与开发,延长晚期肿瘤患者的生存时间。